[Mechanism of polyphyllin â
targeting EGFR to affect proliferation and apoptosis of human breast cancer cells].
Zhongguo Zhong Yao Za Zhi
; 47(3): 721-729, 2022 Feb.
Article
em Zh
| MEDLINE
| ID: mdl-35178955
ABSTRACT
This study aims to investigate the molecular mechanism of polyphyllin â
(PPâ
) inhibiting proliferation of human breast cancer cells. Human breast cancer BT474 and MDA-MB-436 cells were treated with different concentrations of PPâ
, and then the effect of PPâ
on cell proliferation was detected by MTT assay, trypan blue dye exclusion assay, real-time cell analysis, and clone forming assay, respectively. The apoptosis was detected by Annexin V-FITC/PI staining and then analyzed by flow cytometry. The change of mitochondrial membrane potential was detected by flow cytometry after fluorescent probe JC-1 staining. Western blot was used to detect protein expression and phosphorylation. Molecular docking was performed to detect the binding between PPâ
and EGFR. The affinity between PPâ
and EGFR was determined by drug affinity responsive target stability assay. The results indicated that PPâ
inhibited the proliferation and colony formation of BT474 and MDA-MB-436 cells in a time-and concentration-dependent manner. The PPâ
treatment group showed significantly increased apoptosis rate and significantly decreased mitochondrial membrane potential. PPâ
down-regulated the expression of pro-caspase-3 protein, promoted the cleavage of PARP, and significantly reduced the phosphorylation levels of EGFR, Akt, and ERK. Molecular docking showed that PPâ
bound to the extracellular domain of EGFR and formed hydrogen bond with Gln366 residue. Drug affinity responsive target stability assay confirmed that PPâ
significantly prevented pronase from hydrolyzing EGFR, indicating that PPâ
and EGFR have a direct binding effect. In conclusion, PPâ
inhibited the proliferation and induced apoptosis of breast cancer cells by targeting EGFR to block its downstream signaling pathway. This study lays a foundation for the further development of PPâ
-targeted drugs against breast cancer.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Neoplasias da Mama
Limite:
Female
/
Humans
Idioma:
Zh
Revista:
Zhongguo Zhong Yao Za Zhi
Assunto da revista:
FARMACOLOGIA
/
TERAPIAS COMPLEMENTARES
Ano de publicação:
2022
Tipo de documento:
Article
País de afiliação:
China